DOCSLIB.ORG

Vmc 605: Systematic Animal Virology Retroviridae

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Vmc 605: Systematic Animal Virology Retroviridae VMC 605: SYSTEMATIC ANIMAL VIROLOGY RETROVIRIDAE Dr Manoj Kumar Assistant Professor Department of Veterinary Microbiology Bihar Animal Sciences University Retro: from Latin retro,"backwards” - refers to the activity of reverse RETROVIRIDAE transcriptase and the transfer of genetic information from RNA to DNA. Retrovirus: A retrovirus is a lysogenic virus with an RNA genome that uses reverse transcriptase to make DNA for insertion into the host genome. Retroviruses • RNA viruses • single stranded, positive sense, enveloped, icosahedral. • Distinguished from all other RNA viruses by presence of an unusual enzyme, reverse transcriptase. Retroviruses • Retro = reversal • RNA is serving as a template for DNA synthesis. • One genera of veterinary interest • Alpharetrovirus • • Family - Retroviridae • Subfamily - Orthoretrovirinae [Ortho: from Greek orthos"straight" • Genus -. Alpharetrovirus • Genus - Betaretrovirus Family- • Genus - Gammaretrovirus • Genus - Deltaretrovirus Retroviridae • Genus - Lentivirus [ Lenti: from Latin lentus, "slow“ ]. • Genus - Epsilonretrovirus • Subfamily - Spumaretrovirinae • Genus - Spumavirus Retroviridae • Subfamily • Orthoretrovirinae • Genus • Alpharetrovirus Alpharetrovirus • Species • Avian leukosis virus(ALV) • Rous sarcoma virus (RSV) • Avian myeloblastosis virus (AMV) • Fujinami sarcoma virus (FuSV) • ALVs have been divided into 10 envelope subgroups - A , B, C, D, E, F, G, H, I & J based on • host range Avian • receptor interference patterns • neutralization by antibodies leukosis- • subgroup A to E viruses have been divided into two groups sarcoma • Noncytopathic (A, C, and E) • Cytopathic (B and D) virus (ALV) • Cytopathic ALVs can cause a transient cytotoxicity in 30- 40% of the infected cells 1. The viral envelope formed from host cell membrane; contains 72 spiked knobs. 2. These consist of a transmembrane protein TM (gp 41), which is linked to surface protein SU (gp 120) that binds to a cell receptor during infection. 3. The virion has cone-shaped, icosahedral core, Structure containing the major capsid protein 4. Between capsid and envelope is an outer matrix protein, MA 5. Two identical copies of positive sense ssRNA genome (retroviruses are diploid). 6. Enzymes: reverse transcriptase, integrase and protease. Structure - Retrovirus Structure - Alpharetrovirus The viral genome (oncornaviruses). Genome • Virus attaches to host receptors • Fusion with cell membrane. REPLICATION • Internalization and partial uncoating. • ssRNA(+) genome is copied into a linear - NUCLEAR dsDNA molecule by RT • Nuclear entry of the viral dsDNA at the nuclear membrane is disassembled at mitosis. • Viral dsDNA is covalently and randomly integrated into the cell’s genome by the viral integrase (=provirus). • Transcription of provirus by Pol II • Nuclear export of RNAs. • Translation of viral RNAs >>> produces Env, Gag and GagPol polyproteins. • Assembly of the virion at the host cellular membrane and packaging of the viral RNA genome. • Budding through the plasma membrane and release of the virions • Proteolytic processing of the precursors polyproteins by viral protease and maturation of the virions. Replication cycle of • The flow of the early part of the replication cycle goes from receptor Alpharetrovirus binding and internalization at the left through reverse transcription to integration of the proviral DNA. The late part of the replication cycle proceeds from the provirus through transcription • and processing and translation of viral RNA to assembly and release of viral particles. Maturation of the released particles involves cleavage of viral polyproteins by PR (protease). receptor uncoating RNA blocked Replicative receptors viral dsDNA integrated ds DNA into host genome cycle mRNA proteins pro-virus assembly genomic RNA Questions??? Thanks.
Load more

Recommended publications
  • The Expression of Human Endogenous Retroviruses Is Modulated by the Tat Protein of HIV‐1
    The Expression of Human Endogenous Retroviruses is modulated by the Tat protein of HIV‐1 by Marta Jeannette Gonzalez‐Hernandez A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy (Immunology) in The University of Michigan 2012 Doctoral Committee Professor David M. Markovitz, Chair Professor Gary Huffnagle Professor Michael J. Imperiale Associate Professor David J. Miller Assistant Professor Akira Ono Assistant Professor Christiane E. Wobus © Marta Jeannette Gonzalez‐Hernandez 2012 For my family and friends, the most fantastic teachers I have ever had. ii Acknowledgements First, and foremost, I would like to thank David Markovitz for his patience and his scientific and mentoring endeavor. My time in the laboratory has been an honor and a pleasure. Special thanks are also due to all the members of the Markovitz laboratory, past and present. It has been a privilege, and a lot of fun, to work near such excellent scientists and friends. You all have a special place in my heart. I would like to thank all the members of my thesis committee for all the valuable advice, help and jokes whenever needed. Our collaborators from the Bioinformatics Core, particularly James Cavalcoli, Fan Meng, Manhong Dai, Maureen Sartor and Gil Omenn gave generous support, technical expertise and scientific insight to a very important part of this project. Thank you. Thanks also go to Mariana Kaplan’s and Akira Ono’s laboratory for help with experimental designs and for being especially generous with time and reagents. iii Table of Contents Dedication ............................................................................................................................ ii Acknowledgements ............................................................................................................. iii List of Figures ...................................................................................................................
    [Show full text]
  • VMC 321: Systematic Veterinary Virology Retroviridae Retro: from Latin Retro,"Backwards”
    VMC 321: Systematic Veterinary Virology Retroviridae Retro: from Latin retro,"backwards” - refers to the activity of reverse RETROVIRIDAE transcriptase and the transfer of genetic information from RNA to DNA. Retroviruses Viral RNA Viral DNA Viral mRNA, genome (integrated into host genome) Reverse (retro) transfer of genetic information Usually, well adapted to their hosts Endogenous retroviruses • RNA viruses • single stranded, positive sense, enveloped, icosahedral. • Distinguished from all other RNA viruses by presence of an unusual enzyme, reverse transcriptase. Retroviruses • Retro = reversal • RNA is serving as a template for DNA synthesis. • One genera of veterinary interest • Alpharetrovirus • • Family - Retroviridae • Subfamily - Orthoretrovirinae [Ortho: from Greek orthos"straight" • Genus -. Alpharetrovirus • Genus - Betaretrovirus Family- • Genus - Gammaretrovirus • Genus - Deltaretrovirus Retroviridae • Genus - Lentivirus [ Lenti: from Latin lentus, "slow“ ]. • Genus - Epsilonretrovirus • Subfamily - Spumaretrovirinae • Genus - Spumavirus Retroviridae • Subfamily • Orthoretrovirinae • Genus • Alpharetrovirus Alpharetrovirus • Species • Avian leukosis virus(ALV) • Rous sarcoma virus (RSV) • Avian myeloblastosis virus (AMV) • Fujinami sarcoma virus (FuSV) • ALVs have been divided into 10 envelope subgroups - A , B, C, D, E, F, G, H, I & J based on • host range Avian • receptor interference patterns • neutralization by antibodies leukosis- • subgroup A to E viruses have been divided into two groups sarcoma • Noncytopathic (A, C, and E) • Cytopathic (B and D) virus (ALV) • Cytopathic ALVs can cause a transient cytotoxicity in 30- 40% of the infected cells 1. The viral envelope formed from host cell membrane; contains 72 spiked knobs. 2. These consist of a transmembrane protein TM (gp 41), which is linked to surface protein SU (gp 120) that binds to a cell receptor during infection. 3. The virion has cone-shaped, icosahedral core, Structure containing the major capsid protein 4.
    [Show full text]
  • Effects of Retroviruses on Host Genome Function
    ANRV361-GE42-20 ARI 1 August 2008 18:2 V I E E W R S I E N C N A D V A Effects of Retroviruses on Host Genome Function Patric Jern and John M. Coffin Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts 02111; email: [email protected], John.Coffi[email protected] Annu. Rev. Genet. 2008. 42:20.1–20.23 Key Words The Annual Review of Genetics is online at Human Endogenous Retrovirus, LTR, transcription, recombination, genet.annualreviews.org methylation This article’s doi: 10.1146/annurev.genet.42.110807.091501 Abstract Copyright c 2008 by Annual Reviews. For millions of years, retroviral infections have challenged vertebrates, All rights reserved occasionally leading to germline integration and inheritance as ERVs, 0066-4197/08/1201-0001$20.00 genetic parasites whose remnants today constitute some 7% to 8% of the human genome. Although they have had significant evolutionary side effects, it is useful to view ERVs as fossil representatives of retro- viruses extant at the time of their insertion into the germline, not as direct players in the evolutionary process itself. Expression of particu- lar ERVs is associated with several positive physiological functions as well as certain diseases, although their roles in human disease as etio- logical agents, possible contributing factors, or disease markers—well demonstrated in animal models—remain to be established. Here we discuss ERV contributions to host genome structure and function, in- cluding their ability to mediate recombination, and physiological effects on the host transcriptome resulting from their integration, expression, and other events.
    [Show full text]
  • Lentiviral Integration Site Targeting: Host Determinants and Consequences
    University of Pennsylvania ScholarlyCommons Publicly Accessible Penn Dissertations Spring 2010 Lentiviral Integration Site Targeting: Host Determinants and Consequences Keshet Ronen University of Pennsylvania, [email protected] Follow this and additional works at: https://repository.upenn.edu/edissertations Part of the Virology Commons Recommended Citation Ronen, Keshet, "Lentiviral Integration Site Targeting: Host Determinants and Consequences" (2010). Publicly Accessible Penn Dissertations. 174. https://repository.upenn.edu/edissertations/174 This paper is posted at ScholarlyCommons. https://repository.upenn.edu/edissertations/174 For more information, please contact [email protected]. Lentiviral Integration Site Targeting: Host Determinants and Consequences Abstract A necessary step in the retroviral lifecycle is integration, the covalent insertion of the viral cDNA into the genome of the infected cell. This means that retroviruses, for example HIV, establish life-long infection. It also means that retroviruses are used as gene-delivery vectors to treat genetic diseases. Integration events are distributed non-randomly in the genome of the infected cell, with characteristic genus-specific preferences. In this dissertation, we focus on the lentiviral class of retroviruses, and explore two aspects of their integration: the means by which integration is targeted to its favored sites, and the consequences of integration at these sites for the host cell. The host protein LEDGF/p75 has been shown to interact with lentiviral integrases and contribute to their preference for integration in genes. We sought to establish the extent to which integration site selection is determined by LEDGF/p75 tethering. We first asked whether LEDGF/p75 was an essential integration tether, by analyzing integration site distribution in cells stringently depleted for LEDGF/p75.
    [Show full text]
  • TNPO3-Mediated Nuclear Entry of the Rous Sarcoma Virus Gag Protein Is Independent
    bioRxiv preprint doi: https://doi.org/10.1101/2020.03.12.989608; this version posted April 21, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 TNPO3-mediated nuclear entry of the Rous sarcoma virus Gag protein is independent 2 of the cargo-binding domain 3 4 Breanna L. Ricea, Matthew S. Stakea*, and Leslie J. Parenta,b,# 5 6 aDivision of Infectious Diseases and Epidemiology, Department of Medicine, Penn State 7 College of Medicine, Hershey, PA, USA 8 bDepartment of Microbiology and Immunology, Penn State College of Medicine, 9 Hershey, PA, USA 10 11 Running Head: TNPO3-mediated nuclear entry of alpharetrovirus Gag 12 13 #Address correspondence to Leslie Parent, [email protected]. 14 *Present address: 15 Matthew S. Stake 16 UPMC Hanover Medical Group, Hanover, PA, USA 17 18 B.L.R and M.S.S. contributed equally to this work. 19 20 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.03.12.989608; this version posted April 21, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 21 Abstract 22 Retroviral Gag polyproteins orchestrate the assembly and release of nascent 23 virus particles from the plasma membranes of infected cells.
    [Show full text]
  • An Introduction to Viral Vectors: Safety Considerations
    An Introduction to Viral Vectors: Safety Considerations Dawn P. Wooley, Ph.D., SM(NRCM), RBP, CBSP Learning Objectives Recognize hazards associated with viral vectors in research and animal testing laboratories. Interpret viral vector modifications pertinent to risk assessment. Understand the difference between gene delivery vectors and viral research vectors. 2 Outline Introduction to Viral Vectors Retroviral & Lentiviral Vectors (+RNA virus) Adeno and Adeno-Assoc. Vectors (DNA virus) Novel (-)RNA virus vectors NIH Guidelines and Other Resources Conclusions 3 Increased Use of Viral Vectors in Research Difficulties in DNA delivery to mammalian cells <50% with traditional transfection methods Up to ~90% with viral vectors Increased knowledge about viral systems Commercialization has made viral vectors more accessible Many new genes identified and cloned (transgenes) Gene therapy 4 5 6 What is a Viral Vector? Viral Vector: A viral genome with deletions in some or all essential genes and possibly insertion of a transgene Plasmid: Small (~2-20 kbp) circular DNA molecules that replicates in bacterial cells independently of the host cell chromosome 7 Molecular Biology Essentials Flow of genetic information Nucleic acid polarity Infectivity of viral genomes Understanding cDNA cis- vs. trans-acting sequences cis (Latin) – on the same side trans (Latin) – across, over, through 8 Genetic flow & nucleic acid polarity Coding DNA Strand (+) 5' 3' 5' 3' 5' 3' 3' 5' Noncoding DNA Strand (-) mRNA (+) RT 3' 5' cDNA(-) Proteins (Copy DNA aka complementary DNA) 3' 5' 3' 5' 5' 3' mRNA (+) ds DNA in plasmid 9 Virology Essentials Replication-defective vs. infectious virus Helper virus vs. helper plasmids Pathogenesis Original disease Disease caused by transgene Mechanisms of cancer Insertional mutagenesis Transduction 10 Viral Vector Design and Production 1 + Vector Helper Cell 2 + Helper Constructs Vector 3 + + Vector Helper Constructs Note: These viruses are replication-defective but still infectious.
    [Show full text]
  • Animal Models for HIV AIDS: a Comparative Review
    Comparative Medicine Vol 57, No 1 Copyright 2007 February 2007 by the American Association for Laboratory Animal Science Pages 33-43 Animal Models for HIV AIDS: A Comparative Review Debora S Stump and Sue VandeWoude* Human immunodeficiency virus (HIV), the causative agent for acquired immune deficiency syndrome, was described over 25 y ago. Since that time, much progress has been made in characterizing the pathogenesis, etiology, transmission, and disease syndromes resulting from this devastating pathogen. However, despite decades of study by many investigators, basic questions about HIV biology still remain, and an effective prophylactic vaccine has not been developed. This review provides an overview of the viruses related to HIV that have been used in experimental animal models to improve our knowledge of lentiviral disease. Viruses discussed are grouped as causing (1) nonlentiviral immunodeficiency-inducing diseases, (2) naturally occurring pathogenic infections, (3) experimentally induced lentiviral infections, and (4) nonpathogenic lentiviral infections. Each of these model types has provided unique contributions to our understanding of HIV disease; further, a comparative overview of these models both reinforces the unique attributes of each agent and provides a basis for describing elements of lentiviral disease that are similar across mammalian species. Abbreviations: AIDS, acquired immune deficiency syndrome; BIV, bovine immunodeficiency virus; CAEV, caprine arthritis-encephalitis virus; CRPRC, California Regional Primate Research
    [Show full text]
  • 2002.V043.04: to Create Two Subfamilies Within the Family
    February 19, 2002 From the Retroviridae Study Group 2002.V043.04: To create two subfamilies within the family Retroviridae, the first one would contain the six genera as previously defined: Alpharetrovirus, Betaretrovirus, Gammaretrovirus, Deltaretrovirus, Epsilonretrovirus and Lentivirus. The second one would contain a single genus, Spumavirus. 2002.V044.04: to name the first subfamily Orthoretrovirinae 2002.V045.04: to name the second subfamily Spumaretrovirinae. 2002.V046.04: To incorporate the genera Alpharetrovirus, Betaretrovirus, Gammaretrovirus, Deltaretrovirus, Epsilonretrovirus and Lentivirus, currently genera of the family Retroviridae, within the new subfamily Spumaretrovirinae. 2002.V047.04: To incorporate the genus Spumavirus, currently a genus of the family Retroviridae, within the new subfamily Spumaretrovirinae. 2002.V048.04: To designate Simian foamy virus as the new type species of the genus Spumavirus instead of Champazee foamy virus, now a strain of the species Simian foamy virus. 2002.V049.04: To incorporate in the species Simian foamy virus, the new type species of the genus Spumavirus the strains Chimpanzee foamy virus, Simian foamy virus 1 and Simian foamy virus 3, previously listed as species in the Spumavirus genus _______________________________ 2002.V043.04: To create two subfamilies within the family Retroviridae, the first one would contain the six genera as previously defined: Alpharetrovirus, Betaretrovirus, Gammaretrovirus, Deltaretrovirus, Epsilonretrovirus and Lentivirus. The second one would contain a single genus, Spumavirus. 2002.V044.04: to name the first subfamily Orthoretrovirinae 2002.V045.04: to name the second subfamily Spumaretrovirinae. Background: The background of this proposal is as follows: The Retroviridae Study Group had proposed in the past to separate the family Retroviridae into two subfamilies, to be called Orthoretrovirinae and the Spumaretrovirinae.
    [Show full text]
  • Identification of Diverse Full-Length Endogenous Betaretroviruses In
    Hayward et al. Retrovirology 2013, 10:35 http://www.retrovirology.com/content/10/1/35 RESEARCH Open Access Identification of diverse full-length endogenous betaretroviruses in megabats and microbats Joshua A Hayward1,2, Mary Tachedjian3†, Jie Cui4†, Hume Field5, Edward C Holmes4,6, Lin-Fa Wang3,7,8 and Gilda Tachedjian1,2,9* Abstract Background: Betaretroviruses infect a wide range of species including primates, rodents, ruminants, and marsupials. They exist in both endogenous and exogenous forms and are implicated in animal diseases such as lung cancer in sheep, and in human disease, with members of the human endogenous retrovirus-K (HERV-K) group of endogenous betaretroviruses (βERVs) associated with human cancers and autoimmune diseases. To improve our understanding of betaretroviruses in an evolutionarily distinct host species, we characterized βERVs present in the genomes and transcriptomes of mega- and microbats, which are an important reservoir of emerging viruses. Results: A diverse range of full-length βERVs were discovered in mega- and microbat genomes and transcriptomes including the first identified intact endogenous retrovirus in a bat. Our analysis revealed that the genus Betaretrovirus can be divided into eight distinct sub-groups with evidence of cross-species transmission. Betaretroviruses are revealed to be a complex retrovirus group, within which one sub-group has evolved from complex to simple genomic organization through the acquisition of an env gene from the genus Gammaretrovirus. Molecular dating suggests that bats have contended with betaretroviral infections for over 30 million years. Conclusions: Our study reveals that a diverse range of betaretroviruses have circulated in bats for most of their evolutionary history, and cluster with extant betaretroviruses of divergent mammalian lineages suggesting that their distribution may be largely unrestricted by host species barriers.
    [Show full text]
  • Evolutionary Inference from Endogenous Retrovirus Distribution and Diversity
    Evolutionary Inference from Endogenous Retrovirus Distribution and Diversity Robert James Moncreiff Gifford University of London Imperial College of Science, Technology and Medicine Department of Biology Silwood Park A thesis submitted for the degree of Doctor of Philosophy in the year 2002 1 Preface PREFACE The work in this thesis carried out between October 1998 and August 2002. My research was supported by a studentship from the Natural Environment Research Council (NERC) and supervised by Dr M. Tristem. This thesis is the result of my own work except where explicitly stated in the text. The contents have not been previously submitted for any degree, diploma, or any other qualification at Imperial College or at any other university. 2 Acknowledgements ACKNOWLEDGEMENTS I would like to thank my friends and colleagues at Silwood Park for their support and understanding. Equally I would like to thank my friends elsewhere, and my family, for offering some respite from retroviruses and from science in general, and for their encouragement and unaccountable faith in me. Special thanks to Paul-Michael Agapow for his invaluable guidance and supervision in the realm of programming and bioinformatics, and to Vicki and her family for their wonderful hospitality during my stint in post-grant purgatory. Above all I would like to thank my supervisor, Dr Mike Tristem, for his support, guidance, and his remarkable patience and generosity. 3 Abstract ABSTRACT Endogenous retroviruses (ERVs) are the relics of germline infections by ancient retroviruses. ERVs are widespread elements within the genomic DNA of vertebrates, and show great potential as markers of evolutionary processes.
    [Show full text]
  • Molecular Characterization of Natural Recombinant Alvs And
    MOLECULAR CHARACTERIZATION OF NATURAL RECOMBINANT ALVS AND THEIR USE AS VECTORS FOR GENE DELIVERY INTO STABLE CELL LINES by TAYLOR MARCELO CORREA BARBOSA (Under the Direction of Guillermo Zavala) ABSTRACT Avian Leukosis viruses (ALV) previously found as contaminant of Marek’s Disease vaccines were molecularly characterized. All three isolates, named PDRC-1039, PDRC-3246 and PDRC- 32-49, were found to be recombinant viruses. The mosaic viruses mostly consist of endogenous ALV sequences but, the surface protein (gp85) is of exogenous origin and very similar to ALV- A subgroup. Due to their high similarity to each other, the viruses were considered to be from a common source, despite being from different vaccine companies. One of the viruses, PDRC- 1039, was used to construct a novel ALV based retroviral defective vector system for gene delivery, named pBZ system. Eight different vectors were constructed to allow versatility of vector utilization. All vectors are bicistronic with one drug resistant gene, which allow selection of transduced cells. All vectors were tested for transfection and transduction using Green Fluorescence Protein (GFP). The titers achieved with transduction were similar for all vectors. An avian fibroblastoid cell line (DF-1) was used for transduction and expression of GFP. Selected cells were positive for fluorescence during all passages tested, which varies from 5 to 25 depending on the vector design. One of the pBZ vectors, the pBZ3.0, was used to transduce the Infectious Laryngotracheitis Virus (ILTV) glycoprotein I (gI) into DF-1 cells. After transduction and drug selection with G-418, single colonies were tested for ILTV gI expression by immunoflourescence and westernblot assays.
    [Show full text]
  • Tat-Independent Lentivirus Genomes for Vaccination and Host/Pathogen Interaction Studies
    THÈSE Pour obtenir le grade de DOCTEUR DE LA COMMUNAUTÉ UNIVERSITÉ GRENOBLE ALPES Spécialité : Virologie/Microbiologie/Immunologie Arrêté ministériel : 7 août 2006 Présentée par Deepanwita BOSE Thèse dirigée par Dr. Yahia Chebloune préparée au sein du Laboratoire Pathogénèse et Vaccination Lentivirales (PAVAL Lab) dans l'École Doctorale Chimie & Sciences du Vivant (ED218) Tat-independent lentivirus genomes for vaccination and host/pathogen interaction studies. Thèse soutenue publiquement le 26 Janvier 2017, devant le jury composé de : Dr. Giuseppe Bertoni Maitre de conférences, IVI, Bern, Suisse Rapporteur Pr. Francois Villinger Professeur, Director of New Iberia Research Center, USA, Rapporteur Pr. Philippe Sabatier Professeur à l’Université Grenoble Alpes, Président Dr.Yahia Chebloune Research Director, à l’Université Grenoble Alpes, Examinateur Dr. Corinne Ronfort Chargé de recherche, Université Lyon I, Examinateur i “Dedicated to my Mom, Dad and Brother” i Acknowledgements Acknowledgements This thesis in its current form is a result of support, assistance, collaboration, critical inputs and guidance of several people and thus, I would sincerely thank each one of them. Firstly, I would like to express my sincere gratitude to my mentor Dr. Yahia Chebloune for having his faith and confidence in me and accepting me in his research lab for doing a PhD. thesis. I am thankful to him for his support, encouragement and extraordinary guidance all through my graduate work. His insightful discussions and critical comments were an invaluable part in preparation of this thesis. I am grateful for his patience throughout the tribulations, errors, repeats and also the successes in the lab, the corrections in the redaction of the thesis.
    [Show full text]